AU2005300248A1 - Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal - Google Patents
Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal Download PDFInfo
- Publication number
- AU2005300248A1 AU2005300248A1 AU2005300248A AU2005300248A AU2005300248A1 AU 2005300248 A1 AU2005300248 A1 AU 2005300248A1 AU 2005300248 A AU2005300248 A AU 2005300248A AU 2005300248 A AU2005300248 A AU 2005300248A AU 2005300248 A1 AU2005300248 A1 AU 2005300248A1
- Authority
- AU
- Australia
- Prior art keywords
- kit
- vector
- gene
- nucleic acid
- parts according
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000011282 treatment Methods 0.000 title claims description 12
- 230000000259 anti-tumor effect Effects 0.000 title description 6
- 241000124008 Mammalia Species 0.000 title description 3
- 230000000840 anti-viral effect Effects 0.000 title description 2
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 101
- 239000013598 vector Substances 0.000 claims abstract description 86
- 239000003814 drug Substances 0.000 claims abstract description 49
- 229940079593 drug Drugs 0.000 claims abstract description 44
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 28
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 21
- 239000002243 precursor Substances 0.000 claims abstract description 19
- XRECTZIEBJDKEO-UHFFFAOYSA-N flucytosine Chemical compound NC1=NC(=O)NC=C1F XRECTZIEBJDKEO-UHFFFAOYSA-N 0.000 claims description 46
- 230000000694 effects Effects 0.000 claims description 37
- 108010080611 Cytosine Deaminase Proteins 0.000 claims description 31
- 102000000311 Cytosine Deaminase Human genes 0.000 claims description 31
- 206010028980 Neoplasm Diseases 0.000 claims description 30
- 101100502558 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) FCY2 gene Proteins 0.000 claims description 25
- 229920001184 polypeptide Polymers 0.000 claims description 25
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 25
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 25
- 108091000036 uracil phosphoribosyltransferase Proteins 0.000 claims description 24
- 230000003612 virological effect Effects 0.000 claims description 24
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 claims description 23
- 229960002949 fluorouracil Drugs 0.000 claims description 23
- 239000013612 plasmid Substances 0.000 claims description 20
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 241000701161 unidentified adenovirus Species 0.000 claims description 19
- 241000700605 Viruses Species 0.000 claims description 17
- 230000037430 deletion Effects 0.000 claims description 16
- 238000012217 deletion Methods 0.000 claims description 16
- 201000011510 cancer Diseases 0.000 claims description 14
- 229960002963 ganciclovir Drugs 0.000 claims description 13
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 12
- 201000010099 disease Diseases 0.000 claims description 10
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 10
- IRSCQMHQWWYFCW-UHFFFAOYSA-N ganciclovir Chemical compound O=C1NC(N)=NC2=C1N=CN2COC(CO)CO IRSCQMHQWWYFCW-UHFFFAOYSA-N 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 10
- HBOMLICNUCNMMY-XLPZGREQSA-N zidovudine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](N=[N+]=[N-])C1 HBOMLICNUCNMMY-XLPZGREQSA-N 0.000 claims description 9
- 241001430294 unidentified retrovirus Species 0.000 claims description 8
- 101100502554 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) FCY1 gene Proteins 0.000 claims description 7
- 229960004150 aciclovir Drugs 0.000 claims description 6
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 claims description 6
- -1 fludarcibine Chemical compound 0.000 claims description 6
- 239000013603 viral vector Substances 0.000 claims description 6
- HDOVUKNUBWVHOX-QMMMGPOBSA-N Valacyclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCOC(=O)[C@@H](N)C(C)C)C=N2 HDOVUKNUBWVHOX-QMMMGPOBSA-N 0.000 claims description 5
- WREGKURFCTUGRC-POYBYMJQSA-N Zalcitabine Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](CO)CC1 WREGKURFCTUGRC-POYBYMJQSA-N 0.000 claims description 5
- 125000002091 cationic group Chemical group 0.000 claims description 5
- 229940104302 cytosine Drugs 0.000 claims description 5
- ODKNJVUHOIMIIZ-RRKCRQDMSA-N floxuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ODKNJVUHOIMIIZ-RRKCRQDMSA-N 0.000 claims description 5
- 229940093257 valacyclovir Drugs 0.000 claims description 5
- 229960002555 zidovudine Drugs 0.000 claims description 5
- GFFXZLZWLOBBLO-BWVDBABLSA-N 4-amino-1-[(2r,4s,5r)-3-(fluoromethylidene)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one Chemical compound O=C1N=C(N)C=CN1[C@H]1C(=CF)[C@H](O)[C@@H](CO)O1 GFFXZLZWLOBBLO-BWVDBABLSA-N 0.000 claims description 4
- SYMHUEFSSMBHJA-UHFFFAOYSA-N 6-methylpurine Chemical compound CC1=NC=NC2=C1NC=N2 SYMHUEFSSMBHJA-UHFFFAOYSA-N 0.000 claims description 4
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 claims description 4
- 101000807008 Homo sapiens Uracil phosphoribosyltransferase homolog Proteins 0.000 claims description 4
- 101710101148 Probable 6-oxopurine nucleoside phosphorylase Proteins 0.000 claims description 4
- 102000030764 Purine-nucleoside phosphorylase Human genes 0.000 claims description 4
- 102000006601 Thymidine Kinase Human genes 0.000 claims description 4
- 108020004440 Thymidine kinase Proteins 0.000 claims description 4
- 102100037717 Uracil phosphoribosyltransferase homolog Human genes 0.000 claims description 4
- XRMLXZVSFIBRRJ-PEFMBERDSA-N 1-[(1S,3S,4R,5S)-3-hydroxy-4-(hydroxymethyl)bicyclo[3.1.0]hexan-1-yl]thymine Chemical compound O=C1NC(=O)C(C)=CN1[C@@]1(C[C@H](O)[C@H]2CO)[C@H]2C1 XRMLXZVSFIBRRJ-PEFMBERDSA-N 0.000 claims description 3
- WYWHKKSPHMUBEB-UHFFFAOYSA-N 6-Mercaptoguanine Natural products N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 claims description 3
- ODZBBRURCPAEIQ-DJLDLDEBSA-N Brivudine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(C=CBr)=C1 ODZBBRURCPAEIQ-DJLDLDEBSA-N 0.000 claims description 3
- PTOAARAWEBMLNO-KVQBGUIXSA-N Cladribine Chemical compound C1=NC=2C(N)=NC(Cl)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)O1 PTOAARAWEBMLNO-KVQBGUIXSA-N 0.000 claims description 3
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 claims description 3
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 3
- JNTOCHDNEULJHD-UHFFFAOYSA-N Penciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(CCC(CO)CO)C=N2 JNTOCHDNEULJHD-UHFFFAOYSA-N 0.000 claims description 3
- XNKLLVCARDGLGL-JGVFFNPUSA-N Stavudine Chemical compound O=C1NC(=O)C(C)=CN1[C@H]1C=C[C@@H](CO)O1 XNKLLVCARDGLGL-JGVFFNPUSA-N 0.000 claims description 3
- 102100037357 Thymidylate kinase Human genes 0.000 claims description 3
- 241000251539 Vertebrata <Metazoa> Species 0.000 claims description 3
- 229960002436 cladribine Drugs 0.000 claims description 3
- 229960000928 clofarabine Drugs 0.000 claims description 3
- WDDPHFBMKLOVOX-AYQXTPAHSA-N clofarabine Chemical compound C1=NC=2C(N)=NC(Cl)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@@H]1F WDDPHFBMKLOVOX-AYQXTPAHSA-N 0.000 claims description 3
- 229960000684 cytarabine Drugs 0.000 claims description 3
- 108010000742 dTMP kinase Proteins 0.000 claims description 3
- 239000005549 deoxyribonucleoside Substances 0.000 claims description 3
- 229960005277 gemcitabine Drugs 0.000 claims description 3
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 claims description 3
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 3
- 108020004707 nucleic acids Proteins 0.000 claims description 3
- 102000039446 nucleic acids Human genes 0.000 claims description 3
- 229960001179 penciclovir Drugs 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 229950010147 troxacitabine Drugs 0.000 claims description 3
- RXRGZNYSEHTMHC-BQBZGAKWSA-N troxacitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1O[C@@H](CO)OC1 RXRGZNYSEHTMHC-BQBZGAKWSA-N 0.000 claims description 3
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 claims description 2
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 claims description 2
- BXZVVICBKDXVGW-NKWVEPMBSA-N Didanosine Chemical compound O1[C@H](CO)CC[C@@H]1N1C(NC=NC2=O)=C2N=C1 BXZVVICBKDXVGW-NKWVEPMBSA-N 0.000 claims description 2
- 241000714192 Human spumaretrovirus Species 0.000 claims description 2
- 229960004117 capecitabine Drugs 0.000 claims description 2
- 229960002656 didanosine Drugs 0.000 claims description 2
- 229960004396 famciclovir Drugs 0.000 claims description 2
- GGXKWVWZWMLJEH-UHFFFAOYSA-N famcyclovir Chemical compound N1=C(N)N=C2N(CCC(COC(=O)C)COC(C)=O)C=NC2=C1 GGXKWVWZWMLJEH-UHFFFAOYSA-N 0.000 claims description 2
- 229960000961 floxuridine Drugs 0.000 claims description 2
- 108020001507 fusion proteins Proteins 0.000 claims description 2
- 102000037865 fusion proteins Human genes 0.000 claims description 2
- JTEGQNOMFQHVDC-NKWVEPMBSA-N lamivudine Chemical compound O=C1N=C(N)C=CN1[C@H]1O[C@@H](CO)SC1 JTEGQNOMFQHVDC-NKWVEPMBSA-N 0.000 claims description 2
- 229960001627 lamivudine Drugs 0.000 claims description 2
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 claims description 2
- 229960001428 mercaptopurine Drugs 0.000 claims description 2
- 229960003087 tioguanine Drugs 0.000 claims description 2
- 241001529453 unidentified herpesvirus Species 0.000 claims description 2
- 229960000523 zalcitabine Drugs 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims 3
- 108010032220 cyclomaltodextrinase Proteins 0.000 claims 2
- 230000001747 exhibiting effect Effects 0.000 claims 2
- 101150110188 30 gene Proteins 0.000 claims 1
- 241000710929 Alphavirus Species 0.000 claims 1
- 206010010144 Completed suicide Diseases 0.000 claims 1
- 101100428017 Escherichia coli (strain K12) ygjP gene Proteins 0.000 claims 1
- 101100208970 Mus musculus Upp1 gene Proteins 0.000 claims 1
- 206010046865 Vaccinia virus infection Diseases 0.000 claims 1
- 229920006317 cationic polymer Polymers 0.000 claims 1
- MNRILEROXIRVNJ-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=NC=N[C]21 MNRILEROXIRVNJ-UHFFFAOYSA-N 0.000 claims 1
- 101150076274 upp gene Proteins 0.000 claims 1
- 208000007089 vaccinia Diseases 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 78
- 229960004413 flucytosine Drugs 0.000 description 44
- 102000003939 Membrane transport proteins Human genes 0.000 description 19
- 108090000301 Membrane transport proteins Proteins 0.000 description 19
- 238000000034 method Methods 0.000 description 17
- 239000002245 particle Substances 0.000 description 17
- 238000001415 gene therapy Methods 0.000 description 15
- 235000018102 proteins Nutrition 0.000 description 15
- 241000282414 Homo sapiens Species 0.000 description 14
- 239000002773 nucleotide Substances 0.000 description 14
- 125000003729 nucleotide group Chemical group 0.000 description 14
- 230000010076 replication Effects 0.000 description 13
- 241000588724 Escherichia coli Species 0.000 description 11
- 125000003275 alpha amino acid group Chemical group 0.000 description 11
- 230000032258 transport Effects 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 10
- 229940127073 nucleoside analogue Drugs 0.000 description 10
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 8
- 108010078791 Carrier Proteins Proteins 0.000 description 7
- 101150016240 FCY2 gene Proteins 0.000 description 7
- 241001183012 Modified Vaccinia Ankara virus Species 0.000 description 7
- 210000000170 cell membrane Anatomy 0.000 description 7
- 230000002950 deficient Effects 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 229940002612 prodrug Drugs 0.000 description 7
- 239000000651 prodrug Substances 0.000 description 7
- 238000012546 transfer Methods 0.000 description 7
- 108020004684 Internal Ribosome Entry Sites Proteins 0.000 description 6
- 101150021395 JUND gene Proteins 0.000 description 6
- 230000000875 corresponding effect Effects 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 241000701022 Cytomegalovirus Species 0.000 description 5
- 238000010367 cloning Methods 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 230000035945 sensitivity Effects 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 108050008043 Purine-cytosine permeases Proteins 0.000 description 4
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 210000000481 breast Anatomy 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 208000029742 colonic neoplasm Diseases 0.000 description 4
- 238000010276 construction Methods 0.000 description 4
- 230000001472 cytotoxic effect Effects 0.000 description 4
- 101150085096 fur4 gene Proteins 0.000 description 4
- 230000004927 fusion Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 108091006527 nucleoside transporters Proteins 0.000 description 4
- 238000003752 polymerase chain reaction Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 229940035893 uracil Drugs 0.000 description 4
- RNBMPPYRHNWTMA-UAKXSSHOSA-N 5-fluorouridine 5'-monophosphate Chemical compound O1[C@H](COP(O)(O)=O)[C@@H](O)[C@@H](O)[C@@H]1N1C(=O)NC(=O)C(F)=C1 RNBMPPYRHNWTMA-UAKXSSHOSA-N 0.000 description 3
- 206010006187 Breast cancer Diseases 0.000 description 3
- 208000026310 Breast neoplasm Diseases 0.000 description 3
- 102000014778 Concentrative nucleoside transporters Human genes 0.000 description 3
- 108050005111 Concentrative nucleoside transporters Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241001529936 Murinae Species 0.000 description 3
- 241000700618 Vaccinia virus Species 0.000 description 3
- 230000000981 bystander Effects 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000006801 homologous recombination Effects 0.000 description 3
- 238000002744 homologous recombination Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 238000012423 maintenance Methods 0.000 description 3
- 210000004962 mammalian cell Anatomy 0.000 description 3
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000002777 nucleoside Substances 0.000 description 3
- 125000003835 nucleoside group Chemical group 0.000 description 3
- 102000037831 nucleoside transporters Human genes 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 230000000644 propagated effect Effects 0.000 description 3
- 238000007920 subcutaneous administration Methods 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 230000001173 tumoral effect Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- PQGCEDQWHSBAJP-TXICZTDVSA-N 5-O-phosphono-alpha-D-ribofuranosyl diphosphate Chemical compound O[C@H]1[C@@H](O)[C@@H](O[P@](O)(=O)OP(O)(O)=O)O[C@@H]1COP(O)(O)=O PQGCEDQWHSBAJP-TXICZTDVSA-N 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 2
- 229930024421 Adenine Natural products 0.000 description 2
- 241000271566 Aves Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000282465 Canis Species 0.000 description 2
- 102000007644 Colony-Stimulating Factors Human genes 0.000 description 2
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 2
- 241000206602 Eukaryota Species 0.000 description 2
- 101150002048 FUR1 gene Proteins 0.000 description 2
- 241000700588 Human alphaherpesvirus 1 Species 0.000 description 2
- 102100034343 Integrase Human genes 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 108010050904 Interferons Proteins 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 102000000588 Interleukin-2 Human genes 0.000 description 2
- 241000713869 Moloney murine leukemia virus Species 0.000 description 2
- 241000713862 Moloney murine sarcoma virus Species 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 2
- 102000011755 Phosphoglycerate Kinase Human genes 0.000 description 2
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 2
- 241000714474 Rous sarcoma virus Species 0.000 description 2
- 101001059240 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) Site-specific recombinase Flp Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101001099217 Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8) Triosephosphate isomerase Proteins 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 101710117260 Uracil permease Proteins 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229960000643 adenine Drugs 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 101150093170 codA gene Proteins 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 229940047120 colony stimulating factors Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 229960000390 fludarabine Drugs 0.000 description 2
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 210000004897 n-terminal region Anatomy 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 238000001668 nucleic acid synthesis Methods 0.000 description 2
- 238000011580 nude mouse model Methods 0.000 description 2
- 229940118537 p53 inhibitor Drugs 0.000 description 2
- 230000008488 polyadenylation Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 230000000069 prophylactic effect Effects 0.000 description 2
- 150000003230 pyrimidines Chemical class 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 208000037803 restenosis Diseases 0.000 description 2
- 230000001177 retroviral effect Effects 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000011806 swiss nude mouse Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 229940104230 thymidine Drugs 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 102000003390 tumor necrosis factor Human genes 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 210000002845 virion Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DIGQNXIGRZPYDK-WKSCXVIASA-N (2R)-6-amino-2-[[2-[[(2S)-2-[[2-[[(2R)-2-[[(2S)-2-[[(2R,3S)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S,3S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2R)-2-[[2-[[2-[[2-[(2-amino-1-hydroxyethylidene)amino]-3-carboxy-1-hydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxypropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxybutylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-sulfanylpropylidene]amino]-1-hydroxyethylidene]amino]hexanoic acid Chemical compound C[C@@H]([C@@H](C(=N[C@@H](CS)C(=N[C@@H](C)C(=N[C@@H](CO)C(=NCC(=N[C@@H](CCC(=N)O)C(=NC(CS)C(=N[C@H]([C@H](C)O)C(=N[C@H](CS)C(=N[C@H](CO)C(=NCC(=N[C@H](CS)C(=NCC(=N[C@H](CCCCN)C(=O)O)O)O)O)O)O)O)O)O)O)O)O)O)O)N=C([C@H](CS)N=C([C@H](CO)N=C([C@H](CO)N=C([C@H](C)N=C(CN=C([C@H](CO)N=C([C@H](CS)N=C(CN=C(C(CS)N=C(C(CC(=O)O)N=C(CN)O)O)O)O)O)O)O)O)O)O)O)O DIGQNXIGRZPYDK-WKSCXVIASA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 101150096316 5 gene Proteins 0.000 description 1
- VMJCNPSPGKQVDQ-UHFFFAOYSA-N 6-amino-5-fluoro-1H-pyrimidin-2-one 7H-purine Chemical compound N1=CN=C2N=CNC2=C1.FC=1C(=NC(NC1)=O)N VMJCNPSPGKQVDQ-UHFFFAOYSA-N 0.000 description 1
- RJOXFJDOUQJOMQ-UHFFFAOYSA-N 6-sulfanylidene-3,7-dihydropurin-2-one Chemical compound S=C1NC(=O)NC2=C1NC=N2 RJOXFJDOUQJOMQ-UHFFFAOYSA-N 0.000 description 1
- VVIAGPKUTFNRDU-UHFFFAOYSA-N 6S-folinic acid Natural products C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-UHFFFAOYSA-N 0.000 description 1
- 101150065425 7.5K gene Proteins 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 206010059313 Anogenital warts Diseases 0.000 description 1
- 101710081722 Antitrypsin Proteins 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000178270 Canarypox virus Species 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 101150047856 Cav2 gene Proteins 0.000 description 1
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000000907 Condylomata Acuminata Diseases 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 241000724252 Cucumber mosaic virus Species 0.000 description 1
- 108050000190 Cytosine permeases Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 101150059079 EBNA1 gene Proteins 0.000 description 1
- 241000710188 Encephalomyocarditis virus Species 0.000 description 1
- 101150078582 FCY1 gene Proteins 0.000 description 1
- 241000282324 Felis Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000685663 Homo sapiens Sodium/nucleoside cotransporter 1 Proteins 0.000 description 1
- 101000821827 Homo sapiens Sodium/nucleoside cotransporter 2 Proteins 0.000 description 1
- 101000822028 Homo sapiens Solute carrier family 28 member 3 Proteins 0.000 description 1
- 241001135569 Human adenovirus 5 Species 0.000 description 1
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 1
- 102000018251 Hypoxanthine Phosphoribosyltransferase Human genes 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000003814 Interleukin-10 Human genes 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 102000013462 Interleukin-12 Human genes 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- 102000000704 Interleukin-7 Human genes 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 102000003792 Metallothionein Human genes 0.000 description 1
- 108090000157 Metallothionein Proteins 0.000 description 1
- 108010008707 Mucin-1 Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000186366 Mycobacterium bovis Species 0.000 description 1
- 108700019961 Neoplasm Genes Proteins 0.000 description 1
- 102000048850 Neoplasm Genes Human genes 0.000 description 1
- 102000007999 Nuclear Proteins Human genes 0.000 description 1
- 108010089610 Nuclear Proteins Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 241001631646 Papillomaviridae Species 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 108010022233 Plasminogen Activator Inhibitor 1 Proteins 0.000 description 1
- 102000010752 Plasminogen Inactivators Human genes 0.000 description 1
- 108010077971 Plasminogen Inactivators Proteins 0.000 description 1
- 102100039418 Plasminogen activator inhibitor 1 Human genes 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 102100023116 Sodium/nucleoside cotransporter 1 Human genes 0.000 description 1
- 102100021541 Sodium/nucleoside cotransporter 2 Human genes 0.000 description 1
- 102100021470 Solute carrier family 28 member 3 Human genes 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 244000057717 Streptococcus lactis Species 0.000 description 1
- 235000014897 Streptococcus lactis Nutrition 0.000 description 1
- 102000003673 Symporters Human genes 0.000 description 1
- 108090000088 Symporters Proteins 0.000 description 1
- 101150003725 TK gene Proteins 0.000 description 1
- 108010022394 Threonine synthase Proteins 0.000 description 1
- 102000005497 Thymidylate Synthase Human genes 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 208000025009 anogenital human papillomavirus infection Diseases 0.000 description 1
- 201000004201 anogenital venereal wart Diseases 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 239000002259 anti human immunodeficiency virus agent Substances 0.000 description 1
- 230000000340 anti-metabolite Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 230000001475 anti-trypsic effect Effects 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 229940100197 antimetabolite Drugs 0.000 description 1
- 239000002256 antimetabolite Substances 0.000 description 1
- 229940045686 antimetabolites antineoplastic purine analogs Drugs 0.000 description 1
- 229940045688 antineoplastic antimetabolites pyrimidine analogues Drugs 0.000 description 1
- 230000001588 bifunctional effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 108091092328 cellular RNA Proteins 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000009109 curative therapy Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- GYOZYWVXFNDGLU-XLPZGREQSA-N dTMP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)C1 GYOZYWVXFNDGLU-XLPZGREQSA-N 0.000 description 1
- 230000009615 deamination Effects 0.000 description 1
- 238000006481 deamination reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- OZRNSSUDZOLUSN-LBPRGKRZSA-N dihydrofolic acid Chemical compound N=1C=2C(=O)NC(N)=NC=2NCC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OZRNSSUDZOLUSN-LBPRGKRZSA-N 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- VVIAGPKUTFNRDU-ABLWVSNPSA-N folinic acid Chemical compound C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-ABLWVSNPSA-N 0.000 description 1
- 235000008191 folinic acid Nutrition 0.000 description 1
- 239000011672 folinic acid Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 150000002433 hydrophilic molecules Chemical class 0.000 description 1
- 230000003463 hyperproliferative effect Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 231100000568 intoxicate Toxicity 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 210000000867 larynx Anatomy 0.000 description 1
- 229960001691 leucovorin Drugs 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005567 liquid scintillation counting Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- PQLXHQMOHUQAKB-UHFFFAOYSA-N miltefosine Chemical compound CCCCCCCCCCCCCCCCOP([O-])(=O)OCC[N+](C)(C)C PQLXHQMOHUQAKB-UHFFFAOYSA-N 0.000 description 1
- 229960003775 miltefosine Drugs 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 210000005170 neoplastic cell Anatomy 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 238000007474 nonparametric Mann- Whitney U test Methods 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 230000030648 nucleus localization Effects 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 239000002797 plasminogen activator inhibitor Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 125000004219 purine nucleobase group Chemical group 0.000 description 1
- 108010087864 purine permease Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 230000037432 silent mutation Effects 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000001226 triphosphate Substances 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
- C12N15/861—Adenoviral vectors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Oncology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Communicable Diseases (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicinal Preparation (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The present invention relates to a kit of parts comprising a nucleic acid sequence encoding a permease and a drug comprising one nucleobase moiety or a precursor thereof. The present invention further relates to a kit of parts comprising a precursor of a drug comprising a gene coding a permease and a nucleic acid sequence comprising a suicide gene. The present invention also relates to a vector comprising a gene coding a permease and a suicide gene.
Description
WO 2006/048768 PCT/IB2005/003435 1 Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal. The present invention relates to a kit of parts comprising a nucleic 5 acid sequence encoding a permease and a drug comprising one nucleobase moiety or a precursor thereof. The present invention further relates to a kit of parts comprising a precursor of a drug comprising one nucleobase moiety, a nucleic acid sequence comprising a gene coding a permease and a nucleic acid sequence comprising a suicide gene. The 10 present invention also relates to a vector comprising a gene coding a permease and a suicide gene. The present invention is particularly useful in the context of carrying out a gene therapy treatment of proliferative or infectious diseases. Drugs comprising one nucleobase moiety are the most widely used 15 drugs for the treatment of cancer or viral infection. Among them nucleoside analogues such as cytarabine, gemcitabine, fludarabine, cladribine, troxacitabine and clofarabine are currently used in cancer chemotherapy. Nucleoside analogues are antimetabolites which interfere with nucleic acid synthesis. These agents are generally S-phase specific, 20 and can exert cytotoxic activity either by being incorporated and altering the DNA and RNA macromolecules themselves, and/or by interfering with various enzymes involved in nucleic acid synthesis, and/or by modifying the metabolism of physiologic nucleosides. Nucleoside analogues were also among the first compounds shown 25 to be effective against viral infections. For example, the first four anti-HIV drugs to be approved, AZT, ddl, ddC and D4T; were nucleoside analogues. All four of these drugs and other nucleoside analogues are believed to have a similar mechanism of HIV inhibition, in which the nucleosides are progressively phosphorylated to a 5'-triphosphate, which 30 then acts as a chain terminator in a reverse transcriptase (RT) reaction.
WO 2006/048768 PCT/IB2005/003435 2 Nucleobase analogues are also used in the treatment of cancer. For example fluorouracil (5-FU) is a fluorinated pyrimidine that is metabolized intracellulary to its active form, fluorodeoxyuridine monophophate (FdUMP). The active form inhibits DNA synthesis by 5 inhibiting the normal production of thymidine. Fluorouracil is given intravenously to treat colon, rectum, breast, stomach, and pancreas cancers. Fluorouracil is also available in creams and solutions for topical treatment of some skin cancers and genital warts. The development of gene therapy has opened new possibilities for 10 the use of drug comprising one nucleobase moiety. This particular domain of gene therapy which is called "suicide gene therapy" uses genes whose expression products are able to transform an inactive substance (a prodrug) into a cytotoxic substance, thereby giving rise to cell death. The gene encoding the herpes simplex type I virus thymidine 15 kinase (HSV-1 TK) constitutes the prototype of the suicide genes (Caruso et al., 1993, Proc. NatI. Acad. Sci. USA 90, 7024-7028; Culver et al., 1992, Science 256, 1550-1552; Ram et al., 1997, Nat. Med. 3, .1354-1361). While the TK polypeptide is not toxic as such, it catalyzes the transformation of nucleoside analogues such as acyclovir or ganciclovir 20 (GCV). The modified nucleosides are incorporated into the DNA chains which are in the process of elongation, inhibiting cell division as a consequence. A large number of suicide gene/prodrug pairs are currently available. Those which may more specifically be mentioned Escherichia coli (E. Coli) purine nucleoside phosphorylase and 6-methylpurine 25 deoxyribonucleoside (Sorscher et al., 1994, Gene Therapy 1, 223-238), E. coli guanine phosphoribosyl transferase and 6-thioxanthine (Mzoz and Moolten, 1993, Human Gene Therapy 4, 589-595) and cytosine deaminase (CDase) and 5-fluorocytosine (5FC). In human, the transport of drugs comprising one nucleobase moiety 30 across the cell membrane is passive or made by dedicated membraneous WO 2006/048768 PCT/IB2005/003435 3 transporter. For example, most of the nucleoside analogues are hydrophilic molecules and their transport through the cell membrane is regulated by membrane transporter. In mammalian cells, there are two major nucleoside transporter gene families: the equilibrative nucleoside 5 transporters (ENTs) and the concentrative nucleoside transporters (CNTs). The ENTs are facilitated carrier proteins and the CNTs are Na(+) dependent secondary active transporters (Kong et al., 2004, Curr Drug Metab, 5,:63-84) The nucleoside transporters are implicated in resistance to anticancer nucleoside analogues. A single nucleotide polymorphisms 10 (SNPs) in drug transporters may contribute to interindividual variation in response to nucleoside analogues (Damaraju et al., Oncogene. 2003, 22: 7524-36). Therefore, there is need for a new therapeutic strategy which can circumvent a low or non-existent expression of the nucleoside transporters. Moreover, there is also a need to enhance the intracellular 15 concentration of drugs comprising one nucleobase moiety as the intracellular concentration is correlated with the therapeutic efficiency. One of the goals of the present invention is to improve the intracellular concentration of drugs comprising one nucleobase moiety into cells (e.g. tumoral cells or virally infected cells). This goal is achieved by 20 the transfection of the target cells by a nucleic acid sequence comprising a gene coding a permease. The present invention also aims to improve the efficacy of suicide gene therapy, by using in combination a nucleic acid sequence comprising a gene coding a permease and a nucleic acid sequence comprising a 25 suicide gene. This particular use improves the intracellular concentration of the prodrug (i.e. the precursor of a drug comprising a nucleobase moiety) which can be converted by the protein encoded by the suicide gene. This use may also improve the bystander effect by enhancing the transfer of the drug from the cells transfected by the suicide gene to the 30 cells expressing the permease.
WO 2006/048768 PCT/IB2005/003435 4 With this respect, the present invention also allows to use lower amount of drugs or prodrugs and to reduce the side effects induced by such treatments. For this purpose, the present invention provides a kit of parts 5 comprising: - a drug comprising one nucleobase moiety or a precursor thereof, and - a nucleic acid sequence comprising a gene coding a permease. 10 According to another embodiment, the present invention also provides a kit of parts comprising: - a precursor of a drug comprising one nucleobase moiety, and - a nucleic acid sequence comprising a gene coding a permease, and 15 - a nucleic acid sequence comprising a suicide gene. In a preferred embodiment, the kit of parts according to the invention is intended to be used for the treatment of cancer and/or viral diseases in vertebrates and more particularly in humans. As used herein a drug comprising one nucleobase moiety is a 20 molecule which comprise in its structure only one nucleobase moiety. According to a preferred embodiment of the invention, the drug comprising one nucleobase moiety is able to kill cells and more preferably to kill tumoral cells. According to another preferred embodiment, the drug comprising one nucleobase moiety is able to reduce or prevent the 25 replication of a virus in a cell. According to the invention a nucleobase moiety is chosen from the group consisting in Adenine, Thymine, Guanine, Uracil, Cytosine and analogues thereof. According to a preferred embodiment of the invention, a drug 30 comprising one nucleobase moiety is chosen from the group comprising: WO 2006/048768 PCT/IB2005/003435 5 - nucleoside analogues such as cytarabine, gemcitabine, capecitabine, fludarcibine, cladribine, troxacitabine, clofarabine, azidothymidine (AZT), didanosine (ddl), zalcitabine (ddC), stavudine (d4T), fluoromethylene deoxycytidine (FMdC) 5 and lamivudine, floxuridine (FUdR) and - Pyrimidine analogues such as 5-fluorouracil (5-FU), and - Purine analogues such as thioguanine, mercaptopurine, acyclovir, valacyclovir, famcyclovir, ganciclovir. As used herein, a precursor of a drug comprising one nucleobase 10 moiety refers to a molecule which can be converted by one or more enzymatic reaction into a drug comprising one nucleobase moiety. According to a preferred embodiment of the invention, a precursor of a drug comprising one nucleobase moiety is chosen from the group comprising azidothymidine (AZT), Gancyclovir (GCV), 5-Fluorocytosine (5 15 FC), 5-Fluorouracil (5-FU), 6-Methylpurine deoxyribonucleoside, Ganciclovir elaidic acid ester, penciclovir, Acyclovir, Valacyclovir, (E)-5-(2 bromovinyl)-2'-deoxyuridine,zidovudine, 2'-Exo-methanocarbathymidine. As used herein, permease refers to transmembraneous protein involved in the transfer of a drug comprising one nucleobase moiety, or a 20 precursor thereof through the cell membrane. The permease according to the invention notably refers to purine permease, cytosine permease and nucleoside transporters. According to a preferred embodiment of the invention, permease is a purine or a cytosine permease of S. Cerevisiae. The nucleobase 25 transporters of S. cerevisiae consist of the purine-cytosine permease, known as FCY2, and the uracil permease, known as FUR4. According to a more preferred embodiment of the invention, the permease has an amino acid sequence choosen from the group comprising SEQ ID NO:1 and SEQ ID NO:2.
WO 2006/048768 PCT/IB2005/003435 6 The purine-cytosine permease, FCY2 mediates symport of protons and adenine, guanine, hypoxanthine and cytosine across the yeast plasma membrane (Grenson 1969, Jund and Lacroute 1970, Polak and Grenson 1973, Chevallier et al. 1975, Hopkins et al. 1988). FCY2 protein 5 mediates also the transport of 5-fluorocytosine, an analogue of cytosine (Grenson 1969, Jund and Lacroute 1970). FCY2 gene encodes a protein of 533 amino acids (58 kDa) initially predicted to have 10-12 transmembrane-spanning domains (Weber et al. 1990), with nine now favoured (Ferreira et al. 1999). FCY2 exhibits similar affinities for the 10 purine nucleobases and cytosine (Brethes et al. 1992).Uracil uptake into S. cerevisiae is mediated by the uracil permease, FUR4 (Jund and Lacroute 1970, Jund et al. 1977). FUR4 is a uracil-proton symporter (Hopkins et al. 1988) predicted to be a protein of 633 amino acids (71.7 kDa) with 10 transmembrane domains and long cytoplasmic hydrophylic 15 N- and C-terminal tails (Jund et al. 1988, Garnier et al. 1996). FUR4 protein can also mediates the transport of 5-fluorouracil, an analogue of uracil (Jund and Lacroute 1970). With this respect, according to a preferred embodiment, the permease is chosen from the group comprising FCY2 and Fur4 and 20 analogues thereof. Amino acid sequences of FCY2 and Fur4 are notably available in the swissprot database (accession number P17064 and P05316 respectively). Analogues of Fur4 and FCY2 refers to polypeptide having an amino acid sequence which have at least a degree of identity greater than 70%, advantageously greater than 80%, preferably greater 25 than 90%, and most preferably greater than 95% with the amino acid sequence of the parent protein and which retains the ability to transport a drug comprising one nucleobase moiety through the cell membrane According to another preferred embodiment, the permease is chosen from the group comprising hENT1, hENT2, hCNT1, hCNT2 and 30 hCNT3 and analogues thereof. The amino acid sequences of these WO 2006/048768 PCT/IB2005/003435 7 permeases are available to the one skilled in the art, notably in the swissprot database. Analogues of these polypeptides refers to a polypeptide having an amino acid sequence which have at least a degree of identity greater than 70%, advantageously greater than 80%, preferably 5 greater than 90%, and most preferably greater than 95% with the amino acid sequence of the parent polypeptide and which retains the ability to transport a drug comprising one nucleobase moiety through the cell membrane. The one skilled in the art is able to choose the permease which will 10 be associated with the drug or the precursor of the drug comprising one nucleobase moiety. For example, FCY2 and Fur 4 are preferably associated with 5-FC. Suicide gene refers to a gene coding a protein able to convert a precursor of a drug comprising one nucleobase moiety into a drug 15 comprising one nucleobase moiety. Suicide genes comprised but are not limited to genes coding protein having a cytosine deaminase activity, a thymidine kinase activity, an uracil phosphoribosyl transferase activity, a purine nucleoside phosphorylase activity and/or a thymidylate kinase activity. 20 Preferably, the suicide gene comprised in the kit of parts according to the invention codes a proteinable to transform the precursor of the drug comprising one nucleobase moiety comprised in the same kit of parts. Examples of Suicide genes and corresponding precursors of a drug comprising one nucleobase moiety are disclosed in the following table: 25 30 WO 2006/048768 PCT/IB2005/003435 8 Suicide gene precursor of a drug comprising one nucleobase moiety Thymidine Kinase Ganciclovir; Ganciclovir elaidic acid ester; penciclovir; Acyclovir; Valacyclovir; (E)-5-(2-bromovinyl)-2'-deoxyurid ine; zidovudine; 2'-Exo-methanocarbathymidine Cytosine deaminase 5-Fluorocytosine Purine nucleoside phosphorylase 6-Methylpurine deoxyriboside; Fludarabine uracil phosphoribosyl transferase 5-Fluorocytosine; 5-Fluorouracil thymidylate kinase. Azidothymidine According to a preferred embodiment of the invention, the suicide gene according to the invention codes a protein having at least a CDase activity. CDase is involved in the pyrimidine metabolic pathway by which 5 exogenous cytosine is transformed into uracil by means of a hydrolytic deamination. While CDase activities have been demonstrated in prokaryotes and lower eukaryotes (Jund and Lacroute, 1970, J. Bacteriol. 102, 607-615; Beck et al., 1972, J. Bacteriol. 110, 219-228; De Haan et al., 1972, Antonie van Leeuwenhoek 38, 257-263; Hoeprich et al., 1974, J. 10 Inf. Dis. 130, 112-118; Esders and Lynn, 1985, J. Biol. Chem. 260, 3915 3922), they are not present in mammals (Koechlin et al., 1966, Biochem Pharmacol. 15, 435-446; Polak et al., 1976, Chemotherapy 22, 137-153). The Saccharomyces cerevisiae (S. cerevisiae) FCY1 and the E. coli codA genes, which respectively encode the CDase of these two organisms, are WO 2006/048768 PCT/IB2005/003435 9 known and their sequences have been published (EP 402 108; Erbs et al., 1997, Curr. Genet. 31, 1-6; W093/01281). CDase also deaminates an analogue of cytosine, i.e. 5 fluorocytosine (5-FC), thereby forming 5-fluorouracil (5-FU), which is a 5 compound which is highly cytotoxic when it is converted into 5-fluoro-UMP (5-FUMP). Cells which lack CDase activity, either because of a mutation which inactivates the gene encoding the enzyme or because they are naturally deficient in this enzyme, as are mammalian cells, are resistant to 5-FC (Jund and Lacroute, 1970, J. Bacteriol, 102, 607-615; Kilstrup et al., 10 1989, J. Bacteriol. 1989 171, 2124-2127). By contrast, mammalian cells into which the sequences encoding CDase activity were transferred became sensitive to 5-FC (Huber et al., 1993, Cancer Res. 53, 4619-4626; Mullen et al., 1992, Proc. Natl. Acad. Sci. USA 89, 33-37; WO 93/01281). In addition, the neighboring, untransformed cells also become sensitive to 15 5-FC (Huber et al., 1994, Proc. Nati. Acad. Sci. USA 91, 8302-8306). This phenomenon, which is termed a bystander effect, is due to the cells which are expressing the CDase activity secreting 5-FU, which then intoxicates the neighboring cells by straightforward diffusion across the plasma membrane. This property of 5-FU in diffusing passively represents an 20 advantage as compared with the tk/GCV reference system, where the bystander effect requires there to be contact with the cells which are expressing tk (Mesnil et al., 1996, Proc. Nati. Acad. Sci. USA 93, 1831 1835). All the advantages which CDase offers within the context of gene therapy, in particular anticancer gene therapy, can therefore be readily 25 understood. With this respect, according to a more preferred embodiment of the invention, the gene coding a protein having a CDase activity is FCY1 or CodA or an analogue thereof. Analogues of these genes refers to a gene having an nucleic acid sequence which have at least a degree of identity 30 greater than 70%, advantageously greater than 80%, preferably greater WO 2006/048768 PCT/IB2005/003435 10 than 90%, and most preferably greater than 95% with the nucleic acid sequence of the parent gene Patent application FR2004/001657 discloses a gene coding a protein having an improved CDase activity. This polypeptide derives from a native CDase by addition of an amino acid 5 sequence. According to another preferred embodiment of the invention, the protein having a CDase activity is a polypeptide disclosed in FR2004/001657 and more preferably the polypeptide represented in the sequence identifier SEQ ID NO:1 of FR2004/001657. In prokaryotes and lower eukaryotes, uracil is transformed into UMP 10 by the action of uracil phosphoribosyl transferase (UPRTase). This enzyme converts 5-FU into 5-FUMP. According to another preferred embodiment the suicide gene according to the invention code a protein having an UPRTase activity. The UPRTase in question in the present invention may be of any 15 origin, in particular of prokaryotic, fungal or yeast origin. By way of illustration, the nucleic acid sequences encoding the UPRTases from E. coli (Anderson et al., 1992, Eur. J. Biochem 204, 51-56), from Lactococcus lactis (Martinussen and Hammer, 1994, J. Bacteriol. 176, 6457-6463), from Mycobacterium bovis (Kim et al., 1997, Biochem Mol. 20 Biol. Int 41, 1117-1124) and from Bacillus subtilis (Martinussen et al., 1995, J. Bacteriol. 177, 271-274) may be used in the context of the invention. However, it is most particularly preferred to use a yeast UPRTase and in particular that encoded by the S. cerevisiae FUR1 gene whose sequence disclosed in Kern et al. (1990, Gene 88, 149-157) is 25 introduced here by way of reference. As a guide, the sequences of the genes and those of the corresponding UPRTases may be found in the literature .and the specialist databanks (SWISSPROT, EMBL, Genbank, Medline and the like). Moreover, application PCT/FR99/00904 describes an FUR1 gene 30 lacking 105 nucleotides in 5' of the coding part allowing the synthesis of a WO 2006/048768 PCT/IB2005/003435 11 UPRTase from which the 35 first residues have been deleted at the N-terminal position and starting with the methionine at position 36 in the native protein. The product of expression of the mutant gene, designated FURIA105, is capable of complementing an furl mutant of S. cerevisiae. 5 In addition, the truncated mutant exhibits a higher UPRTase activity than that of the native enzyme. Thus, according to a particularly advantageous embodiment, the polypeptide encoded by the suicide gene according to the invention is a deletion mutant of a native UPRTase. The deletion is preferably located in the N-terminal region of the original UPRTase. It may 10 be complete (affecting all the residues of said N-terminal region) or partial (affecting one or more continuous or discontinuous residues in the primary structure). In general, a polypeptide consists of N-terminal, central and C-terminal parts, each representing about a third of the molecule. For example, since the S. cerevisiae UPRTase has 251 amino acids, its 15 N-terminal part consists of the first 83 residues starting with the so-called initiator methionine situated at the first position of the native form. As for the E. coli UPRTase, its N-terminal part covers positions 1 to 69. A preferred protein having an UPRTase activity comprises an amino acid sequence substantially as represented in the sequence 20 identifier SEQ ID NO: 1 of PCT/FR99/00904, starting with the Met residue at position 1 and ending with the Val residue at position 216. The term "substantially" refers to a degree of identity with said sequence SEQ ID NO: I of PCT/FR99/00904 greater than 70%, advantageously greater than 80%, preferably greater than 90%, and most preferably greater than 95%. 25 More preferably still, it comprises the amino acid sequence represented in the sequence identifier SEQ ID NO: 1 of PCT/FR99/00904. As mentioned above, it may comprise additional mutations. There may be mentioned in particular the substitution of the serine residue at position 2 (position 37 in the native UPRTase) by an alanine residue.
WO 2006/048768 PCT/IB2005/003435 12 According to another preferred embodiment, the suicide gene of the invention codes a protein having at least one Cdase and one UPRTase activity. Patent applications W096/16183 and PCT/FR99/00904 describe the use of a fusion protein encoding an enzyme with two domains having 5 the CDase and UPRTase activities and demonstrate that the transfer of a hybrid gene codA::upp or FCY1::FURI or FCYI::FURIA105 carried by an expression plasmid increases the sensitivity of the transfected B16 cells to 5-FC. The protein and nucleic sequences described in these two applications are incorporated into the description of the present 10 application. Although the fusion can take place at any site of the first polypeptide, the N- or C-terminal ends are preferred and in particular the N-terminal end. Advantageously, the fusion in phase uses a second polypeptide having a cytosine deaminase (CDase) activity and which is 15 derived from a native cytosine deaminase, such that the fusion polypeptide according to the invention exhibits the CDase and UPRTase activities. An FCY1::FUR1 fusion is preferred. Such a bifunctional polypeptide makes it possible to improve the sensitivity of the target cells to 5-FC and to 5-FU. 20 A CDase of prokaryotic or lower eukaryotic origin is used. More preferably still, it is a yeast CDase, and in particular that encoded by the Saccharomyces cerevisiae FCYI gene. The cloning and the sequencing of the genes encoding CDases from various sources are available in the literature and specialist databanks. It should be mentioned that the 25 sequence of the FCY1 gene is disclosed in Erbs et al. (1997, Curr. Genet. 31, 1-6). It is of course possible to use a mutant of CDase having a conversion capacity which is comparable to or greater than that of the native enzyme. Persons skilled in the art are capable of cloning the CDase or 30 UPRTase sequences from the published data and of carrying out possible WO 2006/048768 PCT/IB2005/003435 13 mutations, of testing the enzymatic activity of the mutant forms in an acellular or cellular system according to the prior art technology or based on the protocol indicated in application PCT/FR99/00904, and of fusing, in particular in phase, the polypeptides with CDase and UPRTase activity, 5 and consequently all or part of the corresponding genes. According to a more preferred embodiment of the invention, the suicide gene codes a polypeptide comprising an amino acid sequence substantially as represented in the sequence identifier SEQ ID NO: 2 of PCT/FR99/00904, starting with the Met residue at position 1 and ending 10 with the Val residue at position 373. The term "substantially" has the definition given above. A polypeptide comprising the amino acid sequence as represented in the sequence identifier SEQ ID NO: 2 of PCT/FR99/00904 is most particularly suitable for carrying out the invention. 15 Another preferred encoded polypeptide is disclosed in W096/16183 and more preferably the polypeptide encoded by coda::upp or FCY1::FUR1. The nucleic acid sequences may be easily obtained by cloning, by PCR or by chemical synthesis according to the conventional techniques in 20 use. They may be native genes or genes derived from the latter by mutation, deletion, substitution and/or addition of one or more nucleotides. Moreover, their sequences are widely described in the literature which can be consulted by persons skilled in the art. The present invention also relates to a kit of parts as presented 25 above, characterized in that said nucleic acid sequences are inserted into one or more recombinant vectors of plasmid or viral origin, and to such recombinant vectors carrying such nucleotide sequences placed under the control of the elements necessary for their expression in a host cell. More particularly, the kit of parts of the invention may comprise said 30 nucleic acid sequence comprising a gene coding a permease and said WO 2006/048768 PCT/IB2005/003435 14 nucleic acid sequence comprising a suicide gene inserted into the same recombinant vector or into distinct recombinant vectors. According to a preferred embodiment, the nucleic acid sequences are comprised into the same recombinant vector. 5 The present invention also relates to a recombinant vector which carries at least one of the nucleotide acid sequences according to the invention which is placed under the control of the elements which are required for expressing it in a host cell. More particularly, the present invention relates to a recombinant vector comprising a nucleic acid 10 sequence coding a permease. The recombinant vector can be of plasmid or viral origin and can, where appropriate, be combined with one or more substances which improve the transfectional efficiency and/or stability of the vector. These substances are widely documented in the literature which is available to 15 the skilled person (see, for example, Feigner et al., 1987, Proc. West. Pharmacol. Soc. 32, 115-121; Hodgson and Solaiman, 1996, Nature Biotechnology 14, 339-342; Remy et al., 1994, Bioconjugate Chemistry,. 5, 647-654). By way of non-limiting illustration, the substances can be polymers, lipids, in particular cationic lipids, liposomes, nuclear proteins or 20 neutral lipids. These substances can be used alone or in combination. A combination which can be envisaged is that of a recombinant plasmid vector which is combined with cationic lipids (DOGS, DC-CHOL, spermine-chol, spermidine-chol, etc.), lysophospholipides (for example Hexadecylphosphocholine) and neutral lipids (DOPE). 25 According to a preferred embodiment, the cationic lipids which can be used in the present invention are the cationic lipids describes in EP901463B1 and more preferably pcTG90. The choice of the plasmids which can be used within the context of the present invention is immense. They can be cloning vectors and/or 30 expression vectors. In a general manner, they are known to the skilled WO 2006/048768 PCT/IB2005/003435 15 person and, while a number of them are available commercially, it is also possible to construct them or to modify them using the techniques of genetic manipulation. Examples which may be mentioned are the plasmids which are derived from pBR322 (Gibco BRL), pUC (Gibco BRL), 5 pBluescript (Stratagene), pREP4, pCEP4 (Invitrogene) or p Poly (Lathe et al., 1987, Gene 57, 193-201). Preferably, a plasmid which is used in the context of the present invention contains an origin of replication which ensures that replication is initiated in a producer cell and/or a host cell (for example, the ColE1 origin will be chosen for a plasmid which is intended 10 to be produced in E. coli and the oriP/EBNA1 system will be chosen if it desired that the plasmid should be self-replicating in a mammalian host cell, Lupton and Levine, 1985, Mol. Cell. Biol. 5, 2533-2542; Yates et al., Nature 313, 812-815). The plasmid can additionally comprise a selection gene which enables the transfected cells to be selected or identified 15 (complementation of an auxotrophic mutation, gene encoding resistance to an antibiotic, etc.). Naturally, the plasmid can contain additional elements which improve its maintenance and/or its stability in a given cell (cer sequence, which promotes maintenance of a plasmid in monomeric form (Summers and Sherrat, 1984, Cell 36, 1097-1103, sequences for 20 integration into the cell genome). With regard to a viral vector, it is possible to envisage a vector which is derived from a poxvirus (vaccinia virus, in particular MVA, canarypoxvirus, etc.), from an adenovirus, from a retrovirus, from a herpesvirus, from an aiphavirus, from a foamy virus or from an 25 adenovirus-associated virus. It is possible to use replication competent or replication deficient viral vectors. Preference will be given to using a vector which does not integrate. In this respect, adenoviral vectors and MVA are very particularly suitable for implementing the present invention. According to a preferred embodiment, the viral vector according to 30 the invention derives from a Modified Vaccinia Virus Ankara (MVA). MVA WO 2006/048768 PCT/IB2005/003435 16 vectors and methods to produce such vectors are fully described in European patents EP83286 and EP206920, as well as in Mayr et al. (1975, Infection 3, 6-14) and Sutter et Moss (1992, Proc. NatI. Acad. Sci. USA 89, 10847-10851). According to a more preferred embodiment, the 5 nucleotide sequence according to the invention may be inserted in deletion 1, 11, 111, IV, V and VI of the MVA vector and even more preferably in deletion IlIl (Meyer et al., 1991, J. Gen. Virol. 72, 1031-1038; Sutter et al., 1994, Vaccine 12, 1032-1040). Retroviruses have the property of infecting, and in most cases 10 integrating into, dividing cells and in this regard are particularly appropriate for use in relation to cancer. A recombinant retrovirus according to the invention generally contains the LTR sequences, an encapsidation region and the nucleotide sequence according to the invention, which is placed under the control of the retroviral LTR or of an internal promoter such as 15 those described below. The recombinant retrovirus can be derived from a retrovirus of any origin (murine, primate, feline, human, etc.) and in particular from the MOMuLV (Moloney murine leukemia virus), MVS (Murine sarcoma virus) or Friend murine retrovirus (Fb29). It is propagated in an encapsidation cell line which is able to supply in trans the viral 20 polypeptides gag, pol and/or env which are required for constituting a viral particle. Such cell lines are described in the literature (PA317, Psi CRIP GP + Am-12 etc.). The retroviral vector according to the invention can contain modifications, in particular in the LTRs (replacement of the promoter region with a eukaryotic promoter) or the encapsidation region 25 (replacement with a heterologous encapsidation region, for example the VL3O type) (see French applications 94 08300 and 97 05203) Preference will be also given to using an adenoviral vector which lacks all or part of at least one region which is essential for replication and which is selected from the El, E2, E4 and L1-L5 regions in order to avoid 30 the vector being propagated within the host organism or the environment.
WO 2006/048768 PCT/IB2005/003435 17 A deletion of the El region is preferred. However, it can be combined with (an)other modification(s)-/deletion(s) affecting, in particular, all or part of the E2, E4 and/or L1-L5 regions, to the extent that the defective essential functions are complemented in trans by means of a complementing cell 5 line and/or a helper virus. In this respect, it is possible to use second generation vectors of the state of the art (see, for example, international applications WO-A-94/28152 and WO-A-97/04119). By way of illustration, deletion of the major part of the El region and of the E4 transcription unit is very particularly advantageous. For the purpose of increasing the cloning 10 capacities, the adenoviral vector can additionally lack all or part of. the nonessential E3 region. According to another alternative, it is possible to make use of a minimal adenoviral vector which retains the sequences which are essential for encapsidation, namely the 5' and 3' ITRs (Inverted Terminal Repeat), and the encapsidation region. The various adenoviral 15 vectors, and the techniques for preparing them, are known (see, for example, Graham and Prevect, 1991, in Methods in Molecular Biology, Vol 7, p 109-128; Ed: E.J. Murey, The Human Press mc). Furthermore, the origin of the adenoviral vector according to the invention can vary both from the point of view of the species and from the 20 point of view of the serotype. The vector can be derived from the genome of an adenovirus of human or animal (canine, avian, bovine, murine, ovine, porcine, simian, etc.) origin or from a hybrid which comprises adenoviral genome fragments of at least two different origins. More particular mention may be made of the CAV-l or CAV-2 adenoviruses of 25 canine origin, of the DAV adenovirus of avian origin or of the Bad type 3 adenovirus of bovine origin (Zakharchuk et al., Arch. Virol., 1993, 128: 171-176; Spibey and Cavanagh, J. Gen. Virol. 1989, 70: 165-172; Jouvenne et al., Gene, 1987, 60: 21-28; Mittal et al., J. Gen. Virol., 1995, 76: 93-102). However, preference will be given to an adenoviral vector of WO 2006/048768 PCT/IB2005/003435 18 human origin which is preferably derived from a serotype C- adenovirus, in particular a type 2 or 5 serotype C adenovirus. The term "replication-competent" as used herein refers to a viral vector capable of replicating in a host cell in the absence of any trans 5 complementation. In the context of the present invention, this term also encompasses replication-selective or conditionally-replicative adenoviral vectors which are engineered to replicate better or selectively in cancer or hyperproliferative host cells. According to a preferred embodiment of the invention, the 10 replication competent vector is a replication competent adenoviral vector. These replication competent adenoviral vectors are well known by the one skilled in the art. Among these, adenoviral vectors deleted in the El b region coding the 55kD P53 inhibitor, as in the ONYX-015 virus (Bischoff et al, 1996; Heise et al., 2000; WO 94/18992), are particularly preferred. 15 Accordingly, this virus can be used to selectively infect and kill p53 deficient neoplastic cells. A person of ordinary skill in the art can also mutate and disrupt the p53 inhibitor gene in adenovirus 5 or other viruses according to established techniques. Adenoviral vectors deleted in the E1A Rb binding region can also be used in the present invention. For 20 example, Delta24 virus which is a mutant adenovirus carrying a 24 base pair deletion in the E1A region (Fueyo et al., 2000). Delta24 has a deletion in the Rb binding region and does not bind to Rb. Therefore, replication of the mutant virus is inhibited by Rb in a normal cell. However, if Rb is inactivated and the cell becomes neoplastic, Delta24 is no longer inhibited. 25 Instead, the mutant virus replicates efficiently and lyses the Rb-deficient cell. An adenoviral vector according to the present invention can be generated in vitro in Escherichia coli (E. coli) by ligation or homologous recombination (see, for example, international application WO-A 30 96/17070) or else by recombination in a complementing cell line.
WO 2006/048768 PCT/IB2005/003435 19 The elements required for expression consist of all the elements which enable the nucleotide sequence to be transcribed into RNA and the mRNA to be translated into polypeptide. These elements comprise, in particular, a promoter which may be regulable or constitutive. Naturally, 5 the promoter is suited to the chosen vector and the host cell. Examples which may be mentioned are the eukaryotic promoters of the PGK (phosphoglycerate kinase), MT (metallothionein; Mclvor et al., 1987, Mol. Cell Biol. 7, 838-848), a-1 antitrypsin, CFTR, surfactant, immunoglobulin, p-actin (Tabin et al., 1982, Mol. Cell Biol. 2, 426-436) and SRa (Takebe et 10 al., 1988, Mol. Cell Biol. 8, 466-472) genes, the early promoter of the SV40 virus (Simian virus), the LTR of RSV (Rous sarcoma virus), the HSV-l TK promoter, the early promoter of the CMV virus (Cytomegalovirus)., the p7.5K pH5R, pK1 L, p28 and p11 promoters of the vaccinia virus, and the E1A and MLP adenoviral promoters. The promoter 15 can also be a promoter which stimulates expression in a tumor or cancer cell. Particular mention may be made of the promoters of the MUC-1 gene, which is overexpressed in breast and prostate cancers (Chen et al., 1995, J.. Clin. Invest. 96, 2775-2782), of the CEA (standing for carcinoma embryonic antigen) gene, which is overexpressed in colon cancers 20 (Schrewe et al., 1990, Mol. Cell. Biol. 10, 2738-2748) of the tyrosinase gene, which is overexpressed in melanomas (Vile et al., 1993, Cancer Res. 53, 3860-3864), of the ERBB-2 gene, which is overexpressed in breast and pancreatic cancers (Harris et al., 1994, Gene Therapy 1, 170 175) and of the a-fetoprotein gene, which is overexpressed in liver 25 cancers (Kanai et al., 1997, Cancer Res. 57, 461-465). The cytomegalovirus (CMV) early promoter is very particularly preferred. However, when a vector deriving from a Vaccinia Virus (as for example an MVA vector) is used, the promoter of the thymidine kinase 7.5K gene is particularly preferred.
WO 2006/048768 PCT/IB2005/003435 20 The necessary elements can furthermore include additional elements which improve the expression of the nucleotide sequence according to the invention or its maintenance in the host cell. Intron sequences, secretion signal sequences, nuclear localization sequences, 5 internal sites for the reinitiation of translation of IRES type, transcription termination poly A sequences, tripartite leaders and origins of replication may in particular be mentioned. These elements are known to the skilled person. The recombinant vector according to the invention can also 10 comprise one or more additional genes of interest, with it being possible for these genes to be placed under the control of the same regulatory elements (polycistronic cassette) or of independent elements. Genes which may in particular be mentioned are the genes encoding interleukins IL-2, IL-4, IL-7, IL-10 and IL-12, interferons, tumor necrosis factor (TNF), 15 colony stimulating factors (CSF), in particular GM-CSF, and factors acting on angiogenesis (for example PAI-1, standing for plasminogen activator inhibitor). In one particular embodiment, the recombinant vector according to the invention comprises the gene of interest encoding IL-2 or encoding interferon y (INFy). It is also possible to envisage combining the nucleotide 20 sequence according to the invention with other suicide genes such as the HSV-1 TK gene, the ricin gene, the cholera toxin gene, etc. The present invention also relates to a viral particle which comprises a recombinant vector according to the invention. Such a viral particle can be generated from a viral vector using any technique which is 25 conventional in the field of the art. The viral particle is propagated in a complementing cell which is suited to the deficiencies of the vector. With regard to an adenoviral vector, use will, for example, be made of the 293 cell line, which was established using human embryonic kidney cells and which efficiently complements the El function (Graham et al., 1977, J. 30 Gen. Virol. 36, 59-72), of the A549-E1 cell line (Imler et al., 1996, Gene WO 2006/048768 PCT/IB2005/003435 21 Therapy 3, 75-84) or of a cell line which permits double complementation (Yeh et al., 1996, J. Virol. 70, 559-565; Krougliak and Graham, 1995, Human Gene Therapy 6, 1575-1586; Wang et al., 1995 Gene Therapy 2, 775-783; international application WO 97/04119). It is also possible to 5 employ helper viruses to at least partially complement the defective functions. A complementing cell is understood as being a cell which is able to supply in trans the early and/or late factors which are required for encapsidating the viral genome in a viral capsid in order to generate a viral particle which contains the recombinant vector. Said cell may not be able 10 to complement all the defective functions of the vector on its own and, in this case, can be transfected/transduced with a vector/helper virus which supplies the additional functions. The invention also relates to a process for preparing a viral particle, in which process: 15 (i) a recombinant vector according to the invention is introduced into a complementing cell which is able to complement said vector in trans, so as to obtain a transfected complementing cell, (ii) said transfected complementing cell is cultured under conditions which are appropriate for enabling said viral particle to be 20 produced, and (iii) said viral particle is recovered from the cell culture. While the viral particle can of course be recovered from the culture supernatant, it can also be recovered from the cells. One of the commonly employed methods consists in lysing the cells by means of consecutive 25 freezing/thawing cycles in order to collect the virions in the lysis supernatant. The virions can then be amplified and purified using the techniques of the art (chromatographic method, method of ultra centrifugation, in particular through a cesium chloride gradient, etc.). The present invention, also relates to a composition which 30 comprises a kit of parts, a recombinant vector or a viral particle according WO 2006/048768 PCT/IB2005/003435 22 to the invention in combination with a pharmaceutically acceptable excipient A kit of parts according to the invention is more specifically intended for the preventive or curative treatment of diseases by means of gene 5 therapy and is more specifically aimed at proliferative diseases (cancers, tumors, restenosis, etc.) and at diseases of infectious origin, in particular of viral origin (induced by hepatitis B or C viruses, HIV, herpes, retroviruses, etc.). A kit of parts according to the invention can be made conventionally 10 with a view to administering it locally, parenterally or by the digestive route. In particular, a therapeutically effective quantity of the therapeutic or prophylactic agent is combined with a pharmaceutically acceptable excipient. It is possible to envisage a large number of routes of administration. Examples which may be mentioned are the intragastric, 15 subcutaneous, intracardiac, intramuscular, intravenous,= intraperitoneal, intratumor, intranasal, intrapulmonary and intratracheal routes. In the case of these three latter embodiments, it is advantageous for administration to take place by means of an aerosol or by means of instillation. The administration can take place as a single dose or as a dose which is 20 repeated on one or more occasions after a particular time interval. The appropriate route of administration and dosage vary depending on a variety of parameters, for example the individual, the disease to be treated or the gene(s) of interest to be transferred. The preparations based on viral particles according to the invention can be formulated in the form of 25 doses of between 104 and 1014 pfu (plaque-forming units), advantageously 10 5 and 101 pfu, preferably 106 and 102 pfu, more preferably 109 and 1010 when adenoviral particles are used and 106 and 107 when MVA particles are used. As far as the recombinant vector according to the invention is concerned, it is possible to envisage doses comprising from WO 2006/048768 PCT/IB2005/003435 23 0.01 to 100 mg of DNA, preferably from 0.05 to 10 mg, very particularly preferably from 0.5 to 5 mg. The formulation can also include a diluent, an adjuvant or an excipient which is acceptable from the pharmaceutical point of view, as 5 well as solubilizing, stabilizing and preserving agents. In the case of an injectable administration, preference is given to a formulation in an aqueous, non-aqueous or isotonic solution. It can be presented as a single dose or as a multidose, in liquid or dry (powder, lyophilizate, etc.) form which can be reconstituted at the time of use using an appropriate diluent. 10 The formulation can also comprise appropriate quantities of prodrugs. The present invention also relates to the therapeutic or prophylactic use of a kit of parts, of a recombinant vector or of a viral particle according to the invention for preparing a medicament which is intended for treating the human or animal body by gene therapy. According to a first possibility, 15 the medicament can be administered directly in vivo (for example by intravenous injection, into an accessible tumor, into the lungs by means of an aerosol, into the vascular system using an appropriate catheter, etc.). It is also possible to adopt the ex vivo approach, which consists in removing cells from the patient (bone marrow stem cells, peripheral blood 20 lymphocytes, muscle cells, etc.), transfecting or infecting them in vitro in accordance with the techniques of the art and then readministering them to the patient. A preferred use consists in treating or preventing cancers, tumors and diseases which result , from unwanted cell proliferation. Conceivable applications which may be mentioned are cancers of the 25 breast, of the uterus (in particular those induced by papilloma viruses), of the prostate, of the lung, of the bladder, of the liver, of the colon, of the pancreas, of the stomach, of the esophagus, of the larynx, of the central nervous system (e.g. glioblastoma) and of the blood (lymphomas, leukemia, etc.). It can also be used in the context of cardiovascular 30 diseases, for example in order to inhibit or retard the proliferation of the WO 2006/048768 PCT/IB2005/003435 24 smooth muscle cells of the blood vessel wall (restenosis). Finally, in the case of infectious diseases, it is possible to conceive of the medicament being applied to AIDS. The invention also extends to a method for treating diseases by 5 gene therapy, characterized in that a kit of parts, a recombinant vector, a viral particle or a host cell according to the invention is administered to an host organism or cell which is in need of such treatment. The drug comprising one nucleobase moiety or a precursor thereof can be administered in accordance with standard practice (e.g. per os, 10 systematically). By way of illustration, regarding 5-FU, it is possible to use a dose of from 50 to 500 mg/kg/day, with a dose of 200 mg/kg/day being preferred. Within the context of the present invention, the prodrug is administered in accordance with standard practice (e.g. per os, systematically), with the administration taking place prior to, concomitantly 15 with or else subsequent to the administration of a nucleic acid sequence according to the invention. The oral route is preferred. It is possible to administer a single dose of prodrug or doses which are repeated for a time which is sufficiently long to enable the toxic metabolite to be produced within the host organism or cell. 20 Furthermore, the kit of parts or method according to the invention can be combined with one or more substances which potentiate the cytotoxic effect of the drug comprising one nucleobase moiety. Mention may in particular be made of drugs which inhibit the enzymes of the pathway for the de novo biosynthesis of the pyrimidines (for example 25 those mentioned below), drugs such as Leucovorin (Waxman et al., 1982, Eur. J. Cancer Clin. Oncol. 18, 685-692), which, in the presence of the product of the metabolism of 5-FU (5-FdUMP), increases the inhibition of thymidylate synthase, resulting in a decrease in the pool of dTMP, which is required for replication, and finally drugs such as methotrexate (Cadman 30 et al., 1979, Science 250, 1135-1137) which, by inhibiting dihydrofolate WO 2006/048768 PCT/IB2005/003435 25 reductase and increasing the pool of PRPP (phosphoribosylpyrophosphate), brings about an increase in the incorporation of 5-FU into the cellular RNA. The kit of parts or method according to the invention can be 5 combined with one or more substances effective in anticancer therapy. The kit of parts or methods according to the invention can also be use in combination with a radiotherapy. Figure 1 Antitumor activity of the combination of adenovirus and 5 FC at 125 and 50 mg/kg/day on the growth of LoVo colon cancer 10 xenograft in Swiss nude mice. Tumors were established by s.c. implantation of 5x106 cells into nude mice. When the tumors were palpable, the animals were intratumorally inoculated 3 times with Ad empty (A) Ad-FCU1 (B) and Ad-FCY2FCU1 (C), and treated for 3 weeks with per os administrations of 5-FC. Using Ad-empty (A), no differences in 15 growth curves between absence and presence of 5-FC (P>0.05). Using Ad-FCU1 (B), there is a significant difference in the growth curves between presence and absence of 5-FC only at 125 mg/kg/day (P<0.05). Using Ad-FCY2FCU1 (C), there is a significant difference in the growth curves between without 5-FC and systemic 5-FC at 125 and 50 mg/kg/day 20 (P<0.05). Examples: Construction of an adenovirus expressing FCY2 from the El region: 25 Complete yeast purine-cytosine permease gene (FCY2, 1599 bp) was amplified from the plasmid pAJ14 (Bloch et al. 1992) as template using polymerase chain reaction (PCR). Oligonucleotides SEQ ID N' 3: 5, TAACGATCTCGAGCCATGGTGGAAGAGGGAAATAATGTTTACG-3' 30 (OTG14626, corresponding to the 5' end of the FCY2 coding sequence WO 2006/048768 PCT/IB2005/003435 26 but introducing a Kozak consensus sequence replacing Leu in position 2 by a Val and incorporating an Xhol restriction site upstream of the start codon) and SEQ ID No 4 5' CCCTAATCACGCGTTCCTAACGACCGAAGTATTTTAAT-3' (OTG14627, 5 corresponding to the 3' end and inserting a MIul restriction site downstream of the stop codon) were used as primers. The PCR product was digested by Xhol and Mui and subcloned into the corresponding sites of the mammalian expression vector pCIneo (Promega) generating the plasmid pTG15829. Sequences from pTG15829 revealed a silent mutation 10 at nucleotide +1272 (G to A) in the open reading frame of FCY2 gene. From plasmid pTG15829, the Xhol-M/ul fragment containing the FCY2 gene was inserted into the transfer vector pTG13387 cleaved by the same enzymes, generating the transfer vector pTG1 5857. The adenoviral vector Ad-FCY2 (pTG15895) was generated by homologous recombination in the 15 BJ5183 E. coli strain between the fragment Pacl-BstEll of pTG15857 and the vector pTG6624 linearized by Clal. Final construction Ad-FCY2 (pTG15895) contains a deletion in E3 (nucleotides 28592 to 30470), whereas the El region (nucleotides 459 to 3510) was replaced by the expression cassette containing, from 5' to 3', the CMV immediate-early 20 enhancer/promoter, a chimeric human p-globin/IgG intron, the FCY2 gene, and the bGH polyadenylation signal. The adenoviral particles expressing FCY2 (AdTG15895) were generated by transfection in a cell line complementing the El function (for example line PERC6). 25 Construction of an adenovirus expressing a bicistronic unit FCY2 IRES FCU1 from the El region: The Nhel-Notl fragment of the plasmid pTG15829 containing the FCY2 gene was isolated and introduced into the vector pTG14347 (plasmid pTG4369p53FCU1 described in p53FCU1 patent) linearized with 30 Nhel-Notl. The plasmid pTG16055 thus obtained contains, from 5' to 3', WO 2006/048768 PCT/IB2005/003435 27 the FCY2 gene, the IRES (Internal Ribosome Entry Site) sequence of ECMV (encephalomyocarditis virus), and the FCUI gene. From plasmid pTG16055, the Nhel-BamHl fragment was inserted into the transfer vector pTG14799 (plasmid described in FCU1-8 patent) cleaved by the same 5 enzymes, generating the transfer vector pTG1 6066. The adenoviral vector Ad-FCY2FCU1 (pTG16079) was generated by homologous recombination in the BJ5183 E. coli strain between the fragment Pacl-BstEll of pTG 16066 and the vector pTG6624 linearized by C/al. Final construction Ad FCY2FCU1 (pTG16079) contains a deletion in E3 (nucleotides 28592 to 10 30470), whereas the El region (nucleotides 459 to 3510) was replaced by the expression cassette containing, from 5' to 3', the CMV immediate-early enhancer/promoter, a chimeric human p-globin/IgG intron, the FCY2 IRES FCUI bicistronic unit, and the bGH polyadenylation signal. The adenoviral particles expressing FCY2 IRES FCUI (AdTG16079) were generated by 15 transfection in a cell line complementing the El function (for example line PERC6). 5-fluorocytosine uptake assays: The human tumoral cell line A549 (ATCC CCL-185) was infected 20 with Mock, an empty adenovirus (AdTG15149), an adenovirus expressing FCU1 (AdTG14800 described in FCU1-8 patent) or FCY2 (AdTG15895) or FCY2 and FCUI (AdTG16079). The cells (5.106) were infected at MOI 20 and plated on 60-mm plates. After 24 hours, uptake of 5-FC into transfected cells was measured by conventional tracer techniques. The 25 initial rate of 5-FC (10 ptM, room temperature) was measured using a 1 min incubation period and was traced with [ 3 H]5-FC at a concentration of 0.15 pCi/ml. Briefly, 24 hours after infection, cells were washed once with the transport buffer (100 mM NaCl, 2 mM KCI, 1 mM CaCl 2 , 1mM MgCl 2 and 30 10 mM HEPES, pH 7.5). Uptake assays were started by adding 2 ml of WO 2006/048768 PCT/IB2005/003435 28 transport buffer containing 5-FC at 10 pM ([ 3 H]5-FC at 0.15 pCi/ml). At the end of the incubation, extracellular label was removed by three rapid washes with ice-cold transport buffer. The cells were solubilized in 1 ml of 5% SDS for quantification of radioactivity by liquid scintillation counting. 5 Table 1: Uptake of 5-FC into transfected A549 cells: Uptake of 5-FC was measured using transport buffer containing 10 piM of 5-C. Each values represents the average of three independent 10 experiments ± SD. Vector Uptake of 5-FC (pmol/min/5.10 6 cells) Mock 7.97 ± 3.76 Ad-empty 9.74 ± 3.31 (AdTG15149) Ad-FCU1 13.28 ± 5.74 (AdTG14800) Ad-FCY2 38.94 ± 7.61 (AdTG15895) Ad-FCY2FCU1 77 ± 28.7 (AdTG16079) The results presented in Table I demonstrate an increase in 5-FC transport in cells infected by Adenoviruses expressing FCY2 (AdFCY2 and 15 AdFCY2FCU1). Such results indicate that the yeast FCY2, expressed in human cells, encodes a functional 5-FC transporter.
WO 2006/048768 PCT/IB2005/003435 29 In vitro cell sensitivity to 5-FC: Tumor cells infected with the different adenoviruses were compared for their sensitivity toward 5-FC. The human tumor cell line A549 (ATCC CCL-1 85) was infected with Mock, an empty adenovirus (AdTG1 5149), an 5 adenovirus expressing FCUI (AdTG14800 described in FCU1-8 patent) or FCY2 (AdTG15895) or FCY2 and FCUI (AdTG16079). The cells were infected at an MOI of 1 and then cultured in the presence of different concentrations of 5-FC. After 6 days of culture, the viability of the cells was determined using trypan blue. The LD 5 o values for 5-FC listed in Table 2 10 are the means of four counts. In vivo experiments: The objective of this study is to compare the antitumor activity of the recombinant Ad-FCU1 and Ad-FCY2FCU1 in combination with per os 15 administration of 5-fluorocytosine (5-FC) at 125 or 50 mg/kg/day in nude mice bearing LoVo human colon tumors. LoVo cells were injected subcutaneously (s.c.) into the flanks of Swiss nude mice. Each animal was implanted with 5x106 LoVo cells suspended in 100 ml of PBS. When tumors had reached 30-50 mm3, the 20 mice were randomized in a blinded manner and treated with the indicated vectors at dose of 1.1081U. The vectors (suspended in 100 ml of 10mM Tris-HCI, pH7.5, 1 mM MgCI2) were directly injected into the tumor at days 12, 15 and 18 after tumor implantation. From day 12 on, 5-FC was given by oral gavage at 62.5 mg/kg/day (0.6 ml 5-FC 0.25% in water) or at 25 25 mg/kg/day (0.6 ml 5-FC 0.1% in water) twice a day during 3 weeks. Tumor size was measured in two dimensions using calipers. Tumor volume were calculated in mm3 using the formula (p 6)(length x width2). Statistical analyses were performed using the non-parametric Mann-Whitney U test and Statistica 6.1 software (StatSoft, Inc.). A P<0.05 30 was considered to be statistically significant.
WO 2006/048768 PCT/IB2005/003435 30 Using administration of 5 FC at 125 mg/kg/day, injection of Ad FCU1 and Ad-FCY2FCU1 resulted in a statistically significant suppression of tumor growth (Fig.1B,C). This antitumor effect was specific of FCU1/5 5 FC because infection Ad-empty, which does not induce FCU1 activity, did not significantly reduce tumor size in presence of 5-FC (Fig.1A). At a lower dose of 5-FC (50 mg/kg/day), a significant inhibition of tumor growth was observed after delivery of Ad-FCY2FCU1 (Fig.1C), whereas no modification in tumor growth was observed using Ad-FCU1. This result 10 indicates that the combination of FCY2 and FCU1 is significantly more effective than FCU1 alone and suggests that the FCY2FCU1/5-FC system is more efficient than the FCU1/5-FC system in a cancer gene therapy approach. 15 Table 2: LD 50 values for 5-FC of adenovirus-transduced A549 cells: Vector LD'o (pLM) Mock > 1000 Ad-empty > 1000 (AdTG1 5149) Ad-FCU1 50 (AdTG14800) Ad-FCY2 > 1000 (AdTG15895) Ad-FCY2FCU1 20 (AdTG16079) These results demonstrate that coexpression of the FCY2 and FCUI genes from adenoviral vector (AdTG16079) sensitizes the cells to 5 20 FC more strongly than does the expression of the FCU1 gene WO 2006/048768 PCT/IB2005/003435 31 (AdTG14800). This greater sensitivity to 5-FC results from the uptake of 5 FC encoded by the FCY2 gene. Using cells infected with Ad-FCY2 (AdTG15895), no antiproliferative effect was observed, this indicates that the sole administration of FCY2, in absence of cytosine deaminase 5 activity, is insufficient for an effective antitumor therapy protocol.
Claims (26)
1. A kit of parts for the treatment of cancer and/or viral diseases in vertebrates and more particularly in humans comprising: 5 - a drug comprising one nucleobase moiety or a precursor thereof, and - a nucleic acid sequence comprising a gene coding a permease.
2. A kit of parts for the treatment of cancer and/or viral diseases in 10 vertebrates and more particularly in humans comprising: - a precursor of a drug comprising one nucleobase moiety and, - a nucleic acid sequence comprising a gene coding a permease and, - a nucleic acid sequence comprising a suicide gene. 15
3. A kit of parts according to claim I wherein said drug comprising a nucleobase moiety is selected from the group comprising cytarabine, gemcitabine, capecitabine, fludarcibine, cladribine, troxacitabine, clofarabine, azidothymidine (AZT), didanosine (ddl), zalcitabine (ddC), stavudine (d4T), fluoromethylene deoxycytidine (FMdC) and 20 lamivudine, floxuridine (FUdR), 5-fluorouracil (5-FU), thioguanine, mercaptopurine, acyclovir, valacyclovir, famcyclovir and ganciclovir.
4. A kit of parts according to one of claims 1 to 3 , wherein said permease is a purine or a cytosine peremease of S. Cerevisiae. 25
5. A kit of parts according to claim 4, wherein said permease is chosen from the group comprising FCY2 and Fur4 and analogues thereof
6. A kit of parts according to one of claims 1 to 5 wherein said suicide 30 gene is selected from the group comprising genes coding protein WO 2006/048768 PCT/IB2005/003435 33 having a cytosine deaminase activity, a thymidine kinase activity, an uracil phosphoribosyl transferase activity, a purine nucleoside phosphorylase activity and/or a thymidylate kinase activity 5
7. A kit of parts according to one of claims 1 to 6 wherein said suicide gene codes a protein having a cytosine deaminase activity.
8. A kit of parts according to one of claims I to 7 wherein said suicide gene codes a protein having at least a cytosine deaminase activity and 10 an uracil phosphoribosyl transferase activity.
9. A kit of parts according to claim 7, wherein the suicide gene is selected from the group consisting of CodA, upp, FUR1, FCY1 and FUR1A105 15
10. A kit of parts according to claim 8 wherein said protein is a fusion protein in which a first polypeptide exhibiting a UPRTase or CDase activity is fused in phase with at least a second polypeptide, said second polypeptide exhibiting a CDase or UPRTase activity, 20 respectively.
11. A kit of parts according to claim 8, wherein said suicide gene comprised a first nucleic acid sequence selected from the group consisting of upp, FUR1 and FUR1A105, and a second nucleic acid 25 sequence selected from the group consisting of CodA and FCY1.
12. A kit of parts according to one of claims 1 to 11, wherein said precursor of a drug comprising one nucleobase moiety is chosen from the group comprising azidothymidine (AZT), Gancyclovir (GCV), 5 30 Fluorocytosine (5-FC), 5-Fluorouracil (5-FU), 6-Methylpurine WO 2006/048768 PCT/IB2005/003435 34 deoxyribonucleoside, Ganciclovir elaidic acid ester, penciclovir, Acyclovir, Valacyclovir, (E)-5-(2-bromovinyl)-2' deoxyuridine,zidovudine, 2'-Exo-methanocarbathymidine. 5
13. A kit of parts according to one of claims I to 12, wherein said nucleic acid sequence is inserted into a recombinant vector of plasmid or viral origin
14. A kit of parts according to claim 2, wherein said nucleic acid 10 sequences are inserted into the same recombinant vector.
15. A kit of parts according to claim 2, wherein said nucleic acid sequences are inserted into distinct recombinant vectors. 15
16. A recombinant vector comprising a gene coding a permease and a suicide gene.
17. A recombinant vector according to claim 16, characterized in that said vector is selected from the group consisting of plasmid and viral 20 vectors, where appropriate combined with one or more substances which improve(s) the transfectional efficacy and/or the stability of the vector.
18. A recombinant vector according to claim 17, wherein said substance 25 which improve the transfectional efficacy and/or the stability of the vector is selected from the group comprising cationic lipids, cationic polymers, lysophospholipides and polypeptides.
19. A recombinant vector according to claim 17, wherein said vector is a 30 viral vector which is derived from a pox virus, from an adenovirus, from WO 2006/048768 PCT/IB2005/003435 35 a retrovirus, from a herpes virus, from an alphavirus, from a foamyvirus or from an adenovirusassociated virus.
20. A recombinant Vector according to claim 19, wherein said vector 5 derived from a Modified Vaccinia Ankara (MVA) virus.
21. A recombinant Vector according to claim 20, wherein said gene coding a permease and said suicide gene are inserted at a site of a naturally occurring deletion within the MVA genome selected from the 10 group consisting in deletion 1, 11, 111, IV, V and VI.
22. A recombinant vector according to claim 22, wherein said genes are inserted in the site of the naturally occurring deletion is deletion Ill. 15
23. A recombinant vector according to claim 20, wherein said vector is an adenoviral vector which additionally lacks all or part of the non essential E3 region.
24. A pharmaceutical composition comprising a kit of parts according to 20 one of claims 1 to 15 or a recombinant vector according to one of claims 16 to 23.
25. Use of a pharmaceutical composition according to claim 24 for the preparation of a medicament. 25
26. Use of a pharmaceutical composition according to claim 24 for the preparation of a medicament for the treatment of cancer.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP04360101.2 | 2004-11-08 | ||
EP04360101 | 2004-11-08 | ||
US64659505P | 2005-01-26 | 2005-01-26 | |
US60/646,595 | 2005-01-26 | ||
PCT/IB2005/003435 WO2006048768A2 (en) | 2004-11-08 | 2005-10-26 | Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal |
Publications (2)
Publication Number | Publication Date |
---|---|
AU2005300248A1 true AU2005300248A1 (en) | 2006-05-11 |
AU2005300248B2 AU2005300248B2 (en) | 2012-04-26 |
Family
ID=35539415
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2005300248A Ceased AU2005300248B2 (en) | 2004-11-08 | 2005-10-26 | Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal |
Country Status (18)
Country | Link |
---|---|
US (1) | US8470591B2 (en) |
EP (1) | EP1814907B1 (en) |
JP (2) | JP5372374B2 (en) |
KR (1) | KR101134185B1 (en) |
CN (1) | CN101072792B (en) |
AT (1) | ATE421534T1 (en) |
AU (1) | AU2005300248B2 (en) |
CA (1) | CA2584681C (en) |
CY (1) | CY1109850T1 (en) |
DE (1) | DE602005012535D1 (en) |
DK (1) | DK1814907T3 (en) |
ES (1) | ES2320157T3 (en) |
HK (1) | HK1107360A1 (en) |
IL (1) | IL183015A (en) |
MX (1) | MX2007005474A (en) |
PL (1) | PL1814907T3 (en) |
PT (1) | PT1814907E (en) |
WO (1) | WO2006048768A2 (en) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2136631B1 (en) * | 2007-04-14 | 2012-08-15 | Southern Research Institute | Methods for treating neoplasia with combination of chemotherapeutic agents and radiation |
US9687515B2 (en) | 2007-11-19 | 2017-06-27 | Transgene S.A. | Poxviral oncolytic vectors |
SG178254A1 (en) | 2009-08-07 | 2012-03-29 | Transgene Sa | Composition for treating hbv infection |
TW201221063A (en) * | 2010-12-13 | 2012-06-01 | Tai-Song Guo | Enzyme-assisting yeast, enzyme dough, brewed dough and method of brewing bread, baozi, mantou, bagel and unique enzyme thereof |
EP3129037B1 (en) | 2014-04-10 | 2019-11-13 | Transgene | Poxviral oncolytic vectors |
EP3486322B1 (en) * | 2016-07-13 | 2021-09-29 | National University Corporation Tottori University | Method for producing vaccinia virus expressing foreign gene |
WO2018091680A1 (en) | 2016-11-18 | 2018-05-24 | Transgene Sa | Cowpox-based oncolytic vectors |
US20200032249A1 (en) * | 2017-03-14 | 2020-01-30 | Memorial Sloan Kettering Cancer Center | Labeling, isolation, & analysis of rna from rare cell populations |
CN111826393B (en) * | 2020-08-04 | 2022-02-08 | 湖南农业大学 | Gene for simultaneously regulating grain weight and resistance of rice grain type and application of encoded protein thereof |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5856153A (en) * | 1994-11-17 | 1999-01-05 | Cayla | Suicide genes and new associations of pyrimidine nucleobase and nucleoside analogs with new suicide genes for gene therapy of acquired diseases |
US5858707A (en) * | 1995-07-05 | 1999-01-12 | Schering Corporation | Purified mammalian nucleobase permeases; nucleic acids; antibodies |
US6153740A (en) * | 1995-07-07 | 2000-11-28 | University Of Alberta | cDNA encoding nucleoside transporter |
US6749863B1 (en) * | 1997-11-19 | 2004-06-15 | Georgetown University | Targeted liposome gene delivery |
FR2777570A1 (en) * | 1998-04-17 | 1999-10-22 | Transgene Sa | Novel mutant enzyme and related fusion proteins, useful for gene therapy of cancer, by prodrug activation |
US6552005B1 (en) * | 1998-09-29 | 2003-04-22 | Uab Research Foundation | Molecular chemotherapy enhancement of radiotherapy |
FR2794025A1 (en) * | 1999-05-25 | 2000-12-01 | Transgene Sa | COMPOSITION FOR IMPLEMENTING ANTI-TUMOR OR ANTIVIRAL TREATMENT IN A MAMMAL |
-
2005
- 2005-10-26 MX MX2007005474A patent/MX2007005474A/en active IP Right Grant
- 2005-10-26 EP EP05802308A patent/EP1814907B1/en not_active Not-in-force
- 2005-10-26 PL PL05802308T patent/PL1814907T3/en unknown
- 2005-10-26 ES ES05802308T patent/ES2320157T3/en active Active
- 2005-10-26 US US11/667,254 patent/US8470591B2/en not_active Expired - Fee Related
- 2005-10-26 DK DK05802308T patent/DK1814907T3/en active
- 2005-10-26 JP JP2007539650A patent/JP5372374B2/en not_active Expired - Fee Related
- 2005-10-26 DE DE602005012535T patent/DE602005012535D1/en active Active
- 2005-10-26 KR KR1020077012977A patent/KR101134185B1/en not_active IP Right Cessation
- 2005-10-26 AT AT05802308T patent/ATE421534T1/en active
- 2005-10-26 CA CA2584681A patent/CA2584681C/en not_active Expired - Fee Related
- 2005-10-26 HK HK08101248.0A patent/HK1107360A1/en not_active IP Right Cessation
- 2005-10-26 AU AU2005300248A patent/AU2005300248B2/en not_active Ceased
- 2005-10-26 WO PCT/IB2005/003435 patent/WO2006048768A2/en active Application Filing
- 2005-10-26 CN CN2005800376771A patent/CN101072792B/en not_active Expired - Fee Related
- 2005-11-14 PT PT05802308T patent/PT1814907E/en unknown
-
2007
- 2007-05-06 IL IL183015A patent/IL183015A/en not_active IP Right Cessation
-
2009
- 2009-04-15 CY CY20091100450T patent/CY1109850T1/en unknown
-
2013
- 2013-06-05 JP JP2013119206A patent/JP2013209415A/en not_active Withdrawn
Also Published As
Publication number | Publication date |
---|---|
EP1814907A2 (en) | 2007-08-08 |
EP1814907B1 (en) | 2009-01-21 |
CA2584681A1 (en) | 2006-05-11 |
JP5372374B2 (en) | 2013-12-18 |
MX2007005474A (en) | 2007-07-10 |
CN101072792A (en) | 2007-11-14 |
WO2006048768A8 (en) | 2007-06-14 |
DK1814907T3 (en) | 2009-05-18 |
ES2320157T3 (en) | 2009-05-19 |
JP2008519024A (en) | 2008-06-05 |
KR101134185B1 (en) | 2012-04-09 |
KR20070091614A (en) | 2007-09-11 |
IL183015A (en) | 2012-02-29 |
CA2584681C (en) | 2014-05-13 |
JP2013209415A (en) | 2013-10-10 |
PT1814907E (en) | 2009-03-27 |
WO2006048768A2 (en) | 2006-05-11 |
CN101072792B (en) | 2012-09-26 |
HK1107360A1 (en) | 2008-04-03 |
IL183015A0 (en) | 2007-09-20 |
ATE421534T1 (en) | 2009-02-15 |
CY1109850T1 (en) | 2014-09-10 |
US8470591B2 (en) | 2013-06-25 |
DE602005012535D1 (en) | 2009-03-12 |
WO2006048768A3 (en) | 2006-08-03 |
US20090170795A1 (en) | 2009-07-02 |
PL1814907T3 (en) | 2009-07-31 |
AU2005300248B2 (en) | 2012-04-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2005300248B2 (en) | Kit of parts designed for implementing an antitumoral or antiviral treatment in a mammal | |
US8257692B2 (en) | Polypeptide having an improved cytosine deaminase activity | |
US6596533B1 (en) | Mutant having uracil phosphoribosyl transferase activity | |
AU774391B2 (en) | Composition designed for implementing an antitumoral or antiviral treatment in a mammal | |
AU780074B2 (en) | Composition for implementing a cytotoxic, in particular an antitumoral or antiviral, treatment in a mammal | |
MXPA06000853A (en) | Polypeptide having an improved cytosine deaminase activity | |
AU2009251215A1 (en) | Polypeptide having an improved cytosine deaminase activity |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
DA3 | Amendments made section 104 |
Free format text: THE NATURE OF THE AMENDMENT IS: AMEND THE INVENTION TITLE TO READ KIT OF PARTS DESIGNED FOR IMPLEMENTING AN ANTITUMORAL OR ANTIVIRAL TREATMENT IN A MAMMAL |
|
FGA | Letters patent sealed or granted (standard patent) | ||
MK14 | Patent ceased section 143(a) (annual fees not paid) or expired |